Physical Sciences and Mathematics Commons^™

Evaluation Of Vdr-Coactivator Inhibitors Using Biochemical And Cell-Based Assays, Athena Marie Baranowski

Theses and Dissertations

ABSTRACT

EVALUATION OF VDR–COACTIVATOR INHIBITORS USING BIOCHEMICAL AND CELL–BASED ASSAYS

by

Athena Baranowski

The University of Wisconsin–Milwaukee, 2013

Under the Supervision of Dr. Alexander Arnold

The vitamin D receptor (VDR) is a ligand–dependent transcription factor, which belongs to the nuclear receptor superfamily. VDR–mediated gene regulation is governed by coregulators (coactivators and corepressors). VDR coregulator binding inhibitors (CBIs), which were discovered using high throughput screening (HTS), were evaluated using cell–based assays and biochemical assays to determine their ability to inhibit the interaction between VDR and steroid receptor coactivator–2 (SRC–2). Determining their ability to inhibit the VDR–SRC–2 interaction can lead to the …

Applying Computational Methods To Interpret Experimental Results In Tribology And Enantioselective Catalysis, Michael Garvey

Theses and Dissertations

Computational methods are rapidly becoming a mainstay in the field of chemistry. Advances in computational methods (both theory and implementation), increasing availability of computational resources and the advancement of parallel computing are some of the major forces driving this trend.

It is now possible to perform density functional theory (DFT) calculations with chemical accuracy for model systems that can be interrogated experimentally. This allows computational methods to supplement or complement experimental methods. There are even cases where DFT calculations can give insight into processes and interactions that cannot be interrogated directly by current experimental methods.

This work presents several examples …

An Experimental Investigation Characterizing The Tribological Performance Of Natural And Synthetic Biolubricants Composed Of Carboxylic Acids For Energy Conservation And Sustainability, Carlton Jonathan Reeves

Theses and Dissertations

Over the last several decades the lubrication industry has been striving to bring bio-based lubricants known as biolubricants to prominence. The reasons for the increased environmental initiatives are due to depletion of oil reserves, increases in oil price, stringent government regulations on petroleum-based oils, and most importantly, concerns for protecting the environment. With an estimated, 50% of all lubricants entering the environment and much of these being composed of toxic mineral oils, biolubricants have begun to witness a resurgence. This experimental investigation seeks to develop a new class of ecofriendly biolubricants that are less toxic to the environment, derived from …

I. Studies On The Organocatalytic Formation Of Quaternary Stereocenters. Ii. Studies On The Claisen Rearrangement As A Route To Quaternary Stereocenters. Iii. Asymmetric Synthesis Of Aldehydes Bearing Quaternary Carbon Centers Via The Decarboxylative Asymmetric Allylic Alkylation, Eduardo Alberch

Theses and Dissertations

The asymmetric synthesis of all carbon quaternary stereocenters poses a particular challenge due to the steric congestion inherent in the formation of such centers and has been the object of intense research these last 20-30 years. However, the amount of literature for the synthesis of aldehydes bearing quaternary stereocenters via enolate type chemistry is much more limited due to problems associated with the alkylation of such substrates including such types as Cannizzaro and Tischenko related reactions or self aldol condensations. The formation of aldehydes with quaternary stereocenters via use of enolate equivalents such as the DAAA (decarboxylative asymmetric allylic alkylation) …

Zinc Chemical Biology: The Pursuit Of The Intracellular Targets Of Zinquin, Andrew Nowakowski

Theses and Dissertations

Fluorescent sensors have been a main microscopic tools used to understand Zn2+ physiology on a cellular level. The use of the fluorescent Zn2+ sensor Zinquin (ZQ) and its analogues have revealed that transient Zn2+ is a chief component in a variety of biochemical pathways. Yet, little work has been performed to validate the exact targets of Zinquin in a cellular environment. The goals of this investigation are to determine the types of Zinquin reactions that take place in the cell as well as the identities of its cellular targets.

It has been hypothesized that Zinquin reacts with free Zn2+ within …

Studies In Analytical Chemistry And Chemical Education. Part 1: Characterization Of Complex Organics By Raman Spectroscopy And Gas Chromatography. Part 2: Differential Item Functioning On Multiple-Choice General Chemistry Assessments, Lisa Kay Kendhammer

Theses and Dissertations

PART 1: CHARACTERIZATION OF COMPLEX ORGANICS BY RAMAN SPECTROSCOPY AND GAS CHROMATOGRAPHY.

The analytical chemistry component of this thesis focused on instrumentation and methods to address challenges in art conservation, particularly the identification, quantitation, and reactivity of a set of representative varnishes and their degradation products. Methods for characterizing varnishes are of great interest to art conservators to restore art work more accurately. A database was created as a means to identify and quantify the composition of aged varnishes. Fourier Transform (FT)-Raman Spectroscopy was used to study common organic acids found in varnishes. The database included nine short-chain carboxylic acids, …

Probing Bonding And Dynamics At Heterogeneous Adsorbate/Graphene Interfaces, Eric Charles Mattson

Theses and Dissertations

Graphene-based materials are becoming an astoundingly promising choice for many relevant technological and environmental applications. Deriving graphene from the reduction of graphene oxide (GO) is becoming a popular and inexpensive route toward the synthesis of these materials. While the desired product from GO reduction is pristine graphene, defects and residual oxygen functional groups inherited from the parent GO render reduced graphene oxide (RGO) distinct from graphene. In this work, the structure and bonding for GO and RGO is investigated to the end of a working understanding of the composition and properties of these materials. In situ selected area electron diffraction …

Towards The Use Of Time-Resolved X-Ray Crystallography In Mechanistic Studies Of Cytochrome C Nitrite Reductase From Shewanella Oneidensis, Matthew David Youngblut

Theses and Dissertations

A high-yield expression and purification of Shewanella oneidensis cytochrome c nitrite reductase (ccNiR), and its characterization by a variety of methods, notably Laue crystallography, is reported. A key component of the expression system is an artificial ccNiR gene in which the N-terminal signal peptide from the highly expressed S. oneidensis protein "Small Tetra-heme c" replaces the wild-type signal peptide. This gene, inserted into the plasmid pHSG298 and expressed in S. oneidensis TSP-C strain, generated approximately 20 mg crude ccNiR/L culture, compared with 0.5-1 mg/L for untransformed cells. Purified ccNiR exhibited nitrite and hydroxylamine reductase activities comparable to those of E. …

An Optical-Fiber Based Sensor Array For The Simultaneous Analysis Of Zinc And Copper In Aqueous Environments, Steven Kopitzke

Theses and Dissertations

Copper and zinc are elements commonly used in industrial applications, with many of these processes using these elements in aqueous environments. Before the solutions can be discharged into civil or native waterways, waste treatment processes must be undertaken to ensure compliance with federal, state and local guidelines restricting the concentration of ions discharged in solution. While there are methods of analysis currently available to monitor these solutions, each method has disadvantages, be it that they are cost prohibitive, inaccurate, and/or time-consuming. In this work, a new optical fiber-based platform capable of providing fast and accurate results when performing solution analysis …

Functional Organic Nanomaterials, Ryan R. Kohlmeyer

Theses and Dissertations

π-Conjugated polymers have a wide range of applications such as photovoltaics, light-emitting diodes, and sensors. To gain a better understanding of these systems, monodisperse oligomers can be used as a more simplistic model to generate predictive structural and physical properties of corresponding polymers. A divergent/convergent synthetic approach to synthesis of monodisperse π-conjugated oligomers has been developed. These well-defined, thiophene-containing molecular building blocks have been successfully coupled to a ferrocene hinge, which has been found to be highly efficient in the transport of gold atoms using a gold scanning tunneling microscopy tip.

Carbon nanotubes (CNTs) represent a rare class of materials, …

Characterization Of The Distribution, Abundance, And Fate Of Metals In Sediment, April Grant

Theses and Dissertations

This research describes the mobility of heavy metals in river sediment through characterization of metal-sediment association by a sequential extraction procedure. The most abundant isotopes of As, Cd, Cu, Cr, Pb and Zn were measured by Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) in four sediment extraction phases: a) exchangeable, b) carbonate-bound, c) metal-oxide-bound, and d) organic-bound. The measured concentrations were compared to the Wisconsin Department of Natural Resources' "Sediment Quality Guidelines" to determine whether the metals were present at toxic levels. Two reference techniques, Potentiometric Stripping Analysis and Graphite Furnace Atomic Absorption Spectroscopy, were used to validate the concentrations of metals …

Infrared Light Induced Bending Of Liquid Crystalline Elastomer Composite-Silicone Bilayer Films, Maika Moua

Theses and Dissertations

This research centers on infrared (IR) light-induced bending of liquid crystalline elastomers (LCE) composite-silicone bilayer films. Two new developments are presented in this thesis. First, the reversible infrared (IR) induced bending of 0.1% (w/w) single walled nanotube (SWNT)-LCE/silicone bilayer films were successfully prepared and used for fabrication of functioning devices such as folding, grabbing, and crawling structures. Second, the use of adding a filler (absorbs specific wavelength range), such as Dye 1002, into the LCE matrix achieved wavelength selectivity in LCE systems. The 0.2% (wt/wt) Dye 1002-LCE/silicone bilayer films demonstrated bending under a 980 nm laser source but no bending …

High-Throughput Approaches For The Assessment Of Factors Influencing Bioavailability Of Small Molecules In Pre-Clinical Drug Development, Megan Marie Mccallum

Theses and Dissertations

A bioactive molecule must pass many hurdles to be designated as a "good" pharmaceutical lead or hit compound. It should have a significant activity, selectivity, bioavailability, and metabolic half-life. Many factors have been identified that influence the free drug concentration or bioavailability of orally administered drugs in the earliest development stages. In vitro pre-clinical assays have been developed to measure these parameters. The small molecule properties that are investigated here include aqueous solubility, permeability, reactivity (electrophilicity), small molecule-protein binding, and displacement of protein-bound molecules (drug-drug interactions). The development of rapid and miniaturized assays to quantify these factors is presented herein. …

Methods In Metallomics, Proteomics, And Toxicology: Development And Applications Of Laser Ablation Inductively Coupled Plasma Mass Spectrometry And Native Sds-Page, William John Wobig

Theses and Dissertations

Polyacrylamide gel electrophoresis (PAGE) is a bio-analytical method used to separate proteins in solution into an array of individual bands of proteins in a gel matrix. Current PAGE methods, however, have severe limitations in simultaneously maintaining a protein's native structure and association with transition metals while providing adequate resolution. Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS) provides a means to perform trace to ultra-trace level inorganic analysis of solid samples such as dried PAGE gels containing metallo-protein arrays. Current LA-ICP-MS methods involving the analysis of PAGE gels, however, have been limited in their effective use by inadequate limits of …

Anion Effects In The Extraction Of Metal Ions Into Room-Temperature Ionic Liquids (Rtils), Sarah Lynn Garvey

Theses and Dissertations

As part of a broader program directed at the development of improved approaches to the removal of actinides and heat-producing fission products from spent nuclear fuel, the fundamental aspects of the partitioning of selected metal ions between aqueous solution and various room-temperature ionic liquids (ILs) containing a neutral extractant have been investigated.

Prior work has shown that in contrast to the partitioning of alkaline earth cations into conventional solvents in the presence of a crown ether, which is known to proceed via a single pathway (i.e., neutral complex extraction), their extraction into an IL is often a complex, multi-path process …